Mechanism of action of Astragalus membranaceus for treating diabetic foot ulcers based on single-cell RNA sequencing data and network pharmacology

Diabetic foot ulcer is a severe complication of diabetes, characterized by impaired wound healing and immune dysregulation. Although Astragalus membranaceus (AM) has been widely used and reported to exert beneficial effects in diabetic complications, its underlying mechanisms of action in DFU remain incompletely understood. This study integrated single-cell RNA sequencing (scRNA-seq) data from a public bioinformatic cohort (GSE245703; 4 non-diabetic foot ulcer (NFU) and 5 diabetic foot ulcer (DFU) samples) with network pharmacology to explore potential molecular mechanisms by which AM may be involved in DFU pathology. scRNA-seq analysis identified ten major cell types within the DFU microenvironment and revealed significant macrophage heterogeneity. Network pharmacology identified 14 active compounds in AM and their predicted targets, some of which overlapped with macrophage-associated differentially expressed genes. Molecular docking suggested strong binding affinities between selected AM compounds and macrophage-associated hub genes. qPCR validation in a clinical cohort (6 NFU and 9 DFU patients) confirmed differential expression of several candidate hub genes overlapping with predicted AM targets. Collectively, these results provide a single-cell-resolved, systems-level framework that links AM components to macrophage-associated molecular processes in DFU, offering a hypothesis-generating basis for future functional and translational studies.